环肽cGY-8对小鼠急性缺氧脑损伤的保护作用及转录组学分析  

Protective effect and transcriptomic analysis of cyclic peptide cGY-8 on acute hypoxic brain injury in mice

作　　者：马智慧 李雪雁[1] 张梅[2] 孟盼盼 李亮 唐汉琴 轩莹莹 杨玉田 陈克明 MA Zhihui;LI Xueyan;ZHANG Mei;MENG Panpan;LI Liang;TANG Hanqin;XUAN Yingying;YANG Yutian;CHEN Keming(School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China;Basic Medical Laboratory,No.940 Hospital of Joint Logistics Support Force of Chinese PLA,Lanzhou University of Technology,Lanzhou 730050,China;Key Laboratory of Stem Cells and Gene Drugs of Gansu Province,Lanzhou University of Technology,Lanzhou 730050,China)

机构地区：[1]兰州理工大学生命科学与工程学院,甘肃兰州730050 [2]兰州理工大学解放军联勤保障部队第九四○医院基础医学实验室,甘肃兰州730050 [3]兰州理工大学甘肃省干细胞与基因药物重点实验室,甘肃兰州730050

出　　处：《空军军医大学学报》2024年第12期1412-1419,共8页Journal of Air Force Medical University

基　　金：甘肃省自然科学基金实验动物专项(23JRRA536)。

摘　　要：目的探讨环肽GYHHQYTG(cGY-8)对小鼠急性缺氧脑损伤的保护作用以及其可能的发生机制。方法将BALB/c小鼠随机分为对照组和不同剂量cGY-8给药组,观察各组在常压密闭缺氧条件下小鼠存活时间,筛选出最佳药物浓度。随后将BALB/c小鼠随机分为常氧组(Nor组),模型组(M组),乙酰唑胺组(ACZ组),低、中、高剂量cGY-8给药组,每组6只,使用低压低氧处理后观察各组小鼠脑损伤情况。将15只BALB/c小鼠随机分为常氧组(Nor组)、缺氧组(Hy组)和缺氧给药组(cGY-8组),采用常压密闭缺氧处理后取全脑组织提取RNA进行转录组测序,筛选出差异表达基因(DEGs)后进行GO富集分析和KEGG通路富集分析,并通过qRT-PCR验证测序数据的准确性。结果cGY-8延长小鼠常压缺氧条件下的存活时间,且使低压低氧条件下小鼠脑组织含水量减少、氧化应激反应减轻。转录组学分析发现Hy组与Nor组相比共有678个DEGs(变化大于2倍;P<0.05),而cGY-8处理可回调91个DEGs。GO富集分析91个DEGs主要聚类在信号转导、炎症反应、氧化还原酶活性等;KEGG富集分析可得91个DEGs主要富集在cGMP-PKG信号通路、钙离子信号通路等。结论cGY-8可能通过调节能量代谢、减轻脑细胞Ca^(2+)超载等途径保护小鼠缺氧脑损伤。本研究为预防及治疗高原脑水肿的药物开发提供了新思路。Objective To investigate the protective effect of cyclic peptide GYHHQYTG(cGY-8)on acute hypoxic brain injury in mice and its possible mechanism.Methods BALB/c mice were randomly divided into control group and cGY-8 administration group with different doses to observe the survival time of mice in each group under atmospheric pressure and hypoxia and select the optimal drug concentration.Subsequently,BALB/c mice were randomly divided into normal oxygen group(Nor group),model group(M group),acetazolamide group(ACZ group),and low-dose,medium-dose,and high-dose cGY-8 administration groups,with 6 mice in each group.The brain injury of the mice in each group was observed after hypobaric hypoxia treatment.Fifteen BALB/c mice were randomly divided into normal oxygen group(Nor group),hypoxic group(Hy group),and hypoxic administration group(cGY-8 group).After normal pressure closed hypoxia treatment,RNA was extracted from whole brain tissue for transcripome sequencing,and differentially expressed genes(DEGs)were screened for GO enrichment analysis and KEGG pathway enrichment analysis.The accuracy of the sequencing data was verified by qRT-PCR.Results cGY-8 prolonged the survival time of mice under normal pressure and hypoxia,and reduced the water content and oxidative stress response of mice under low pressure and hypoxia.Transcriptomic analysis showed that there were 678 DEGs in the Hy group compared with the Nor group(the change was more than 2 times;P<0.05),and the cGY-8 treatment could bring back 91 DEGs.GO enrichment analysis showed that 91 DEGs were mainly enriched in signal transduction,inflammatory response,redox enzyme activity,etc.KEGG enrichment analysis showed that 91 DEGs were mainly enriched in cGMP-PKG signal pathway and calcium ion signal pathway.Conclusion cGY-8 may protect mice from hypoxic brain injury by regulating energy metabolism and alleviating Ca^(2+)overload in brain cells.This study provides a new idea for the development of drugs for the prevention and treatment of high-altitude cerebral edema.

关 键 词：环肽cGY-8 急性缺氧脑损伤 RNA-SEQ 小鼠 

分 类 号：R965[医药卫生—药理学]